作　　者： ; ; ;

机构地区： 中国农业大学生物学院农业生物技术国家重点实验室

出　　处： 《农业生物技术学报》 2003年第3期273-275,共3页

摘　　要： 鸡(Gallusgallus)胞外脂肪酸结合蛋白(extracelluarfattyacidbindingprotein,EX-FABP)基因型与鸡腹脂量的积累密切相关。尝试了用等位基因特异性PCR(allele-SpecificPCR,AS-PCR)技术检测EX-FABP基因型的方法,确定了检测的参数和方案,对AS-PCR检验单碱基突变的方法和策略进行了讨论,并在此基础上对该技术进行了改进,建立了等位基因特异性片段长度差异PCR(allele-specificandlength-differentPCR,ASLDPCR)技术。 The genotypes of chick (Gallus gallus ) extracellular fatty acid binding protein (EX-FABP) are closely related to chick abdominal fat percentage. In the study, the method for identifying the EX-FABP genotypes with allele-specific PCR (AS-PCR) technique was established and the strategy of identifying single nucleotide polymorphisms with AS-PCR was discussed. A typical AS-PCR system consists of two allele-specific primers and ane common primer. PCR products can be resulted when a sample's DNA is amplified by the common primer and allele-specific primer which matches with the sequence of sample's DNA. There are not PCR products if the allele-specific primer does not match the sequence of the samples' DNA. According to whether there is PCR products or not, the genotypes of samples can be identified. For establishing an AS-PCR, it is very important to design proper number and kinds of mismatched bases at the 3′ end of the allele-specific primers where the mutation occurs and different genotypes are generated. When one mismatched base in the primers can not lead to different PCR results which can identify genotypes, the second or even the third mismatched bases should be added to the allele-specific primers. The different kinds of mismatched bases have different effect on the results of AS-PCR. If the mismatched bases are A-C or T-G, they have little effect on the amplifying efficiency of AS-PCR. But when the mismatched bases are A-G or C-C or C-G, they have dramatically negative effect on it. A modified AS-PCR, allele-specific and length-deferent PCR (ASLD-PCR) were established and introduced. In an ASLD-PCR system, two pairs of primer sites, each of them consists of an allele specific primer and a common primer,were used to amplify and identify genotypes. The PCR products of the two pairs of primers were different in the length and the different length of PCR products standed for different kinds of genotypes. So ASLD-PCR was more convenient to identify genotypes than AS-PCR.

关 键 词： 等位基因特异性 鸡胞外脂肪酸结合蛋白基因 单碱基突变 鸡

领　　域： [农业科学] [农业科学] [生物学]